Organic compounds

ABSTRACT

A solid pharmaceutical composition suitable for oral administration, comprising: (a) S 1 P receptor agonist; and (b) a sugar alcohol.

The present invention relates to pharmaceutical compositions comprisinga sphingosine-1 phosphate receptor agonist. Sphingosine-1 phosphate(hereinafter “S1P”) is a natural serum lipid. Presently there are 8known S1P receptors, namely S1P1 to S1P8. S1P receptor agonists haveaccelerating lymphocyte homing properties.

S1P receptor agonists are immunomodulating compounds which elicit alymphopenia resulting from a re-distribution, preferably reversible, oflymphocytes from circulation to secondary lymphatic tissue, evoking ageneralized immunosuppression. Naive cells are sequestered, CD4 and CD8T-cells and B-cells from the blood are stimulated to migrate into lymphnodes (LN) and Peyer's patches (PP), and thus infiltration of cells intotransplanted organs is inhibited.

The various known S1P receptor agonists show structural similarities,which result in related problems in providing a suitable formulation. Inparticular, there is a need for an S1P receptor agonist containingformulation which is well-adapted for oral administration in a solidform, e.g. as a tablet or capsule.

Accordingly, the present Invention provides a solid pharmaceuticalcomposition suitable for oral administration, comprising a S1P receptoragonist and a sugar alcohol.

It has surprisingly been found that solid compositions comprising asugar alcohol provide formulations which are particularly well suited tothe oral administration of S1P receptor agonists. The compositionsprovide a convenient means of systemic administration of S1P receptoragonists, do not suffer from the disadvantages of liquid formulationsfor injection or oral use, and have good physicochemical and storageproperties. In particular, the compositions of the present invention mayshow a high level of uniformity in the distribution of the S1P receptoragonist throughout the composition, as well as high stability. Thecompositions of the invention may be manufactured on high speedautomated equipment, and thus do not require hand encapsulation.

S1P receptor agonists are typically sphingosine analogues, such as2-substituted 2-amino-propane-1,3-diol or 2-amino-propanol derivatives.Examples of appropriate S1P receptor agonists are, for example:

-   -   Compounds as disclosed in EP627406A1, e.g. a compound of formula        I

wherein R₁ is a straight- or branched (C₁₂₋₂₂)carbon chain

-   -   which may have in the chain a bond or a hetero atom selected        from a double bond, a triple bond, O, S, NR₆, wherein R₆ is H,        alkyl, aralkyl, acyl or alkoxycarbonyl, and carbonyl, and/or        -   which may have as a substituent alkoxy, alkenyloxy,            alkynyloxy, aralkyloxy, acyl, alkylamino, alkylthio,            acylamino, alkoxycarbonyl, alkoxycarbonylamino, acyloxy,            alkylcarbamoyl, nitro, halogen, amino, hydroxyimino, hydroxy            or carboxy; or

R₁ is

-   -   a phenylalkyl wherein alkyl is a straight- or branched        (C₆₋₂₀)carbon chain; or    -   a phenylalkyl wherein alkyl is a straight- or branched        (C₁₋₃₀)carbon chain wherein said phenylalkyl is substituted by    -   a straight- or branched (C₆₋₂₀)carbon chain optionally        substituted by halogen,    -   a straight- or branched (C₆₋₂₀)alkoxy chain optionally        substituted by halogen,    -   a straight- or branched (C₆₋₂₀)alkenyloxy,    -   phenylalkoxy, halophenylalkoxy, phenylalkoxyalkyl, phenoxyalkoxy        or phenoxyalkyl,    -   cycloalkylalkyl substituted by C₆₋₂₀alkyl,    -   heteroarylalkyl substituted by C₆₋₂₀alkyl,    -   heterocyclic C₆₋₂₀alkyl or    -   heterocyclic alkyl substituted by C₂₋₂₀alkyl,        and wherein        the alkyl moiety may have    -   in the carbon chain, a bond or a heteroatom selected from a        double bond, a triple bond, O, S, sulfinyl, sulfonyl, or NR₆,        wherein R₆ is as defined above, and    -   as a substituent alkoxy, alkenyloxy, alkynyloxy, aralkyloxy,        acyl, alkylamino, alkylthio, acylamino, alkoxycarbonyl,        alkoxycarbonylamino, acyloxy, alkylcarbamoyl, nitro, halogen,        amino, hydroxy or carboxy, and        each of R₂, R₃, R₄ and R₅, independently, is H, C₁₋₄ alkyl or        acyl        or a pharmaceutically acceptable salt thereof;    -   Compounds as disclosed in EP 1002792A1, e.g. a compound of        formula II

wherein m is 1 to 9 and each of R′₂, R′₃, R′₄ and R′₆, independently, isH, alkyl or acyl,or a pharmaceutically acceptable salt thereof;

-   -   Compounds as disclosed in EP0778263 A1, e.g. a compound of        formula III

wherein W is H; C₂₋₆alkenyl or C₂₋₆alkynyl; unsubstituted or by OHsubstituted phenyl; R″₄O(CH₂)_(n); or C₁₋₆alkyl substituted by 1 to 3substituents selected from the group consisting of halogen,C₃₋₈cycloalkyl, phenyl and phenyl substituted by OH;X is H or unsubstituted or substituted straight chain alkyl having anumber p of carbon atoms or unsubstituted or substituted straight chainalkoxy having a number (p-1) of carbon atoms, e.g. substituted by 1 to 3substitutents selected from the group consisting of C₁₋₆ alkyl, OH,acyloxy, amino, C₁₋₆alkylamino, acylamino, oxo, haloC₁₋₆alkyl, halogen,unsubstituted phenyl and phenyl substituted by 1 to 3 substituentsselected from the group consisting of C₁₋₆alkyl, OH, C₁₋₆alkoxy, acyl,acyloxy, amino, C₁₋₆alkylamino, acylamino, haloC₁₋₆alkyl and halogen; Yis H, C₁₋₆alkyl, OH, C₁₋₆alkoxy, acyl, acyloxy, amino, C₁₋₆alkylamino,acylamino, haloC₁₋₆alkyl or halogen, Z₂ is a single bond or a straightchain alkylene having a number or carbon atoms of q,each of p and q, independently, is an integer of 1 to 20, with theproviso of 6≦p+q≦23, m′ is 1, 2 or 3, n is 2 or 3,each of R″₁, R″₂, R″₃ and R″₄, independently, is H, C₁₋₄alkyl or acyl,or a pharmaceutically acceptable salt thereof,

-   -   Compounds as disclosed in WO02/16395, e.g. a compound of formula        IVa or IVb

wherein X_(a) is O, S, NR_(1s) or a group —(CH₂)_(na)—, which group isunsubstituted or substituted by 1 to 4 halogen; n_(a) is 1 or 2, R_(1s)is H or (C₁₋₄)alkyl, which alkyl is unsubstituted or substituted byhalogen; R_(1a) is H, OH, (C₁₋₄)alkyl or O(C₁₋₄)alkyl wherein alkyl isunsubstituted or substituted by 1 to 3 halogen; R_(1b) is H, OH or(C₁₋₄)alkyl, wherein alkyl is unsubstituted or substituted by halogen;each R_(2a) is independently selected from H or (C₁₋₄)alkyl, which alkylis unsubstituted or substituted by halogen; R_(3a) is H, OH, halogen orO(C₁₋₄)alkyl wherein alkyl is unsubstituted or substituted by halogen;and R_(3b) is H, OH, halogen, (C₁₋₄)alkyl wherein alkyl is unsubstitutedor substituted by hydroxy, or O(C₁₋₄)alkyl wherein alkyl isunsubstituted or substituted by halogen; Y_(a) is —CH₂—, —C(O)—,—CH(OH)—, —C(═NOH)—, O or S, and R_(4a) is (C₄₋₁₄)alkyl or(C₄₋₁₄)alkenyl;or a pharmaceutically acceptable salt or hydrate thereof;

-   -   Compounds as disclosed in WO 02/076995, e.g. a compound of        formula V

wherein

-   m_(c) is 1, 2 or 3;-   X_(c) is O or a direct bond;-   R_(1c) is H; C₁₋₆ alkyl optionally substituted by OH, acyl, halogen,    C₃₋₁₀cycloalkyl, phenyl or hydroxy-phenylene; C₂₋₆alkenyl;    C₂₋₆alkynyl; or phenyl optionally substituted by OH;-   R_(2c) is

-    wherein R_(5c) is H or C₁₋₄alkyl optionally substituted by 1, 2 or    3 halogen atoms, and R_(6c) is H or C₁₋₄alkyl optionally substituted    by halogen;-   each of R_(3c) and R_(4c), independently, is H, C₁₋₄alkyl optionally    substituted by halogen, or acyl, and-   R_(c) is C₁₃₋₂₀alkyl which may optionally have in the chain an    oxygen atom and which may optionally be substituted by nitro,    halogen, amino, hydroxy or carboxy; or a residue of formula (a)

-    wherein R_(7c) is H, C₁₋₄alkyl or C₁₋₄alkoxy, and R_(8c) is    substituted C₁₋₂₀alkanoyl, phenylC₁₋₁₄alkyl wherein the C₁₋₁₄alkyl    is optionally substituted by halogen or OH, cycloalkylC₁₋₁₄alkoxy or    phenylC₁₋₁₄alkoxy wherein the cycloalkyl or phenyl ring is    optionally substituted by halogen, C₁₋₄alkyl and/or C₁₋₄alkoxy,    phenylC₁₋₁₄alkoxy-C₁₋₁₄alkyl, phenoxyC₁₋₁₄alkoxy or    phenoxyC₁₋₁₄alkyl,-   R_(c) being also a residue of formula (a) wherein R_(8c) is    C₁₋₁₄alkoxy when R_(1c) is C₁₋₄alkyl, C₂₋₆alkenyl or C₂₋₆alkynyl,    or a compound of formula VI

wherein

-   n_(x) is 2, 3 or 4-   R_(1x) is H; C₁₋₆alkyl optionally substituted by OH, acyl, halogen,    cycloalkyl, phenyl or hydroxy-phenylene; C₂₋₆alkenyl; C₂₋₆alkynyl;    or phenyl optionally substituted by OH;-   R_(2x) is H, C₁₋₄ alkyl or acyl-   each of R_(3x) and R_(4x), independently is H, C₁₋₄alkyl optionally    substituted by halogen or acyl,-   R_(5x) is H, C₁₋₄alkyl or C₁₋₄alkoxy, and-   R_(6x) is C₁₋₂₀ alkanoyl substituted by cycloalkyl;    cyloalkylC₁₋₁₄alkoxy wherein the cycloalkyl ring is optionally    substituted by halogen, C₁₋₄alkyl and/or C₁₋₄alkoxy;    phenylC₁₋₁₄alkoxy wherein the phenyl ring is optionally substituted    by halogen, C₁₋₄alkyl and/or C₁₋₄alkoxy,-   R_(6x) being also C₄₋₁₄alkoxy when R_(1x) is C₂₋₄alkyl substituted    by OH, or pentyloxy or hexyloxy when R_(1x) is C₁₋₄alkyl,    provided that R_(6x) is other than phenyl-butylenoxy when either    R_(5x) is H or R_(1x) is methyl,    or a pharmaceutically acceptable salt thereof;    -   Compounds as disclosed in WO02/06268A1, e.g. a compound of        formula VII

wherein each of R_(1d) and R_(2d), independently, is H or anamino-protecting group;

-   R_(3d) is hydrogen, a hydroxy-protecting group or a residue of    formula

-   R_(4d) is lower alkyl;-   n_(d) is an integer of 1 to 6;-   X_(d) is ethylene, vinylene, ethynylene, a group having a formula    -D-CH₂— (wherein D is carbonyl, —CH(OH)—, O, S or N), aryl or aryl    substituted by up to three substitutents selected from group a as    defined hereinafter,-   Y_(d) is single bond, C₁₋₁₀alkylene, C₁₋₁₀alkylene which is    substituted by up to three substitutents selected from groups a and    b, C₁₋₁₀alkylene having O or S in the middle or end of the carbon    chain, or C₁₋₁₀alkylene having O or S in the middle or end of the    carbon chain which is substituted by up to three substituents    selected from groups a and b;-   R_(5d) is hydrogen, cycloalkyl, aryl, heterocycle, cycloalkyl    substituted by up to three substituents selected from groups a and    b, aryl substituted by up to three substituents selected from groups    a and b, or heterocycle substituted by up to three substituents    selected from groups a and b;-   each of R_(6d) and R_(7d), independently, is H or a substituent    selected from group a;-   each of R_(6d) and R_(9d), independently, is H or C₁₋₄alkyl    optionally substituted by halogen;-   <group a> is halogen, lower alkyl, halogeno lower alkyl, lower    alkoxy, lower alkylthio, carboxyl, lower alkoxycarbonyl, hydroxy,    lower aliphatic acyl, amino, mono-lower alkylamino, di-lower    alkylamino, lower aliphatic acylamino, cyano or nitro; and-   <group b> is cycloalkyl, aryl, heterocycle, each being optionally    substituted by up to three substituents selected from group a;    with the proviso that when R_(5d) is hydrogen, Y_(d) is a group    exclusive of single bond and linear C₁₋₁₀ alkylene, or a    pharmacologically acceptable salt or ester thereof;    -   Compounds as disclosed in JP-14316985 (JP2002316985), e.g. a        compound of formula VIII:

wherein R_(1e), R_(2e), R_(3e), R_(4e), R_(5e), R_(6e), R_(7e), n_(e),X_(e) and Y_(e) are as disclosed in JP-14316985;or a pharmacologically acceptable salt or ester thereof;

-   -   Compounds as disclosed in WO 03/29184 and WO 03/29205, e.g.        compounds of formula IX

wherein X_(f) is O or S, and R_(1f), R_(2f), R_(3f) and n_(f) are asdisclosed in WO 03/29184 and 03/29205, each of R_(4f) and R_(5f),independently is H or a residue of formula

wherein each of R_(8f) and R_(9f), independently, is H or C₁₋₄alkyloptionally substituted by halogen; e.g.2-amino-2-[4-(3-benzyloxyphenoxy)-2-chlorophenyl]propyl-4,3-propane-diolor2-amino-2-[4-(benzyloxyphenylthio)-2-chlorophenyl]propyl-1,3-propane-diol,or a pharmacological salt thereof.

-   -   Compounds as disclosed in WO03/062252A1, e.g. a compound of        formula X

wherein

-   Ar is phenyl or naphthyl; each of mg and ng independently is 0 or 1;    A is selected from COOH, PO3H2, PO2H, SO3H, PO(C1-3alkyl)OH and    1H-tetrazol-5-yl; each of R1g and R2g independently is H, halogen,    OH, COOH or C1-4alkyl optionally substituted by halogen; R3g is H or    C1-4alkyl optionally substituted by halogen or OH; each R4g    independently is halogen, or optionally halogen substituted    C1-4alkyl or C1-3alkoxy; and each of Rg and M has one of the    significances as indicated for B and C, respectively, in    WO03/062252A1;    -   Compounds as disclosed in WO 03/062248A2, e.g. a compound of        formula XI

wherein Ar is phenyl or naphthyl; n is 2, 3 or 4; A is COOH,1H-tetrazol-5-yl, PO3H2, PO2H2, —SO3H or PO(R5h)OH wherein R5h isselected from C1-4alkyl, hydroxyC1-4alkyl, phenyl, —CO—C1-3alkoxy and—CH(OH)-phenyl wherein said phenyl or phenyl moiety is optionallysubstituted; each of R1h and R2h independently is H, halogen, OH, COOH,or optionally halogeno substituted C1-6alkyl or phenyl; R3h is H orC1-4alkyl optionally substituted by halogen and/ OH; each R4hindependently is halogeno, OH, COOH, C1-4alkyl, S(O)0, 1 or 2C1-3alkyl,C1-3alkoxy, C3-6cycloalkoxy, aryl or aralkoxy, wherein the alkylportions may optionally be substituted by 1-3 halogens; and each of Rgand M has one of the significances as indicated for B and C,respectively, in WO03/062248A2.

According to a further embodiment of the invention, a S1P receptoragonist for use in a combination of the invention may also be aselective S1P1 receptor, e.g. a compound which possesses a selectivityfor the S1P1 receptor over the S1P3 receptor of at least 20 fold, e.g.100, 500, 1000 or 2000 fold, as measured by the ratio of EC50 for theS1P1 receptor to the EC50 for the S1P3 receptor as evaluated in a35S-GTPγS binding assay, said compound having an EC50 for binding to theS1P1 receptor of 100 nM or less as evaluated by the 35S-GTPγS bindingassay. Representative S1P1 receptor agonists are e.g. the compoundslisted in WO 03/061567, the contents of which being incorporated hereinby reference, for instance a compound of formula

In each case where citations of patent applications are given, thesubject matter relating to the compounds is hereby incorporated into thepresent application by reference.

Acyl may be a residue R_(y)—CO— wherein R_(y) is C₁₋₆alkyl,C₃₋₆cycloalkyl, phenyl or phenyl-C₁₋₄alkyl. Unless otherwise stated,alkyl, alkoxy, alkenyl or alkynyl may be straight or branched.

When in the compounds of formula I the carbon chain as R₁ issubstituted, it is preferably substituted by halogen, nitro, amino,hydroxy or carboxy. When the carbon chain is interrupted by anoptionally substituted phenylene, the carbon chain is preferablyunsubstituted. When the phenylene moiety is substituted, it ispreferably substituted by halogen, nitro, amino, methoxy, hydroxy orcarboxy.

Preferred compounds of formula I are those wherein R₁ is C₁₃₋₂₀alkyl,optionally substituted by nitro, halogen, amino, hydroxy or carboxy,and, more preferably those wherein R₁ is phenylalkyl substituted byC₆₋₁₄-alkyl chain optionally substituted by halogen and the alkyl moietyis a C₁₋₆alkyl optionally substituted by hydroxy. More preferably, R₁ isphenyl-C₁₋₆alkyl substituted on the phenyl by a straight or branched,preferably straight, C₆₋₁₄alkyl chain. The C₆₋₁₄alkyl chain may be inortho, meta or para, preferably in para.

Preferably each of R₂ to R₅ is H.

A preferred compound of formula I is2-amino-2-tetradecyl-1,3-propanediol. A particularly preferred S1Preceptor agonist of formula I is FTY720, i.e.2-amino-2-[2-(4-octylphenyl)ethyl]propane-1,3-diol in free form or in apharmaceutically acceptable salt form (referred to hereinafter asCompound A), e.g. the hydrochloride, as shown:

A preferred compound of formula If is the one wherein each of R′₂ to R′₅is H and m is 4, i.e.2-amino-2-{2-[4-(1-oxo-5-phenylpentyl)phenyl]ethyl}propane-1,3-diol, infree form or in pharmaceutically acceptable salt form (referred tohereinafter as Compound B), e.g the hydrochloride.

A preferred compound of formula III is the one wherein W is CH₃, each ofR″₁ to R″₃ is H, Z₂ is ethylene, X is heptyloxy and Y is H, i.e.2-amino-4-(4-heptyloxy phenyl)-2-methyl-butanol, in free form or inpharmaceutically acceptable salt form (referred to hereinafter asCompound C), e.g. the hydrochloride. The R-enantiomer is particularlypreferred.

A preferred compound of formula IVa is the FTY720-phosphate (R_(2a) isH, R_(3a) is OH, X_(a) is O, R_(1a) and R_(1b) are OH). A preferredcompound of formula IVb is the Compound C-phosphate (R_(2a) is H, R_(3b)is OH, X_(a) is O, R_(1a) and R_(1b) are OH, Y_(a) is O and R_(4a) isheptyl), A preferred compound of formula V is Compound B-phosphate.

A preferred compound of formula V is phosphoric acidmono-[(R)-2-amino-2-methyl-4-(4-pentyloxy-phenyl)-butyl]ester.

A preferred compound of formula VIII is(2R)-2-amino-4-[3-(4-cyclohexyloxybutyl)-benzo[b]thien-6-yl]-2-methylbutan-1-ol.

When the compounds of formulae I to XIII have one or more asymmetriccenters in the molecule, the various optical isomers, as well asracemates, diastereoisomers and mixtures thereof are embraced.

Examples of pharmaceutically acceptable salts of the compounds offormulae I to XIII include salts with inorganic acids, such ashydrochloride, hydrobromide and sulfate, salts with organic acids, suchas acetate, fumarate, maleate, benzoate, citrate, malate,methanesulfonate and benzenesulfonate salts, or, when appropriate, saltswith metals, such as sodium, potassium, calcium and aluminium, saltswith amines, such as triethylamine and salts with dibasic amino acids,such as lysine. The compounds and salts of the present inventionencompass hydrate and solvate forms.

Binding to S1P receptors can be determined according to the followingassays.

A. Binding Affinity of S1P Receptor Agonists to Individual Human S1PReceptors

Transient Transfection of Human S1P Receptors into HEK293 Cells

S1P receptors and G_(i) proteins are cloned, and equal amounts of 4cDNAs for the EDG receptor, G_(i)-α, G_(i)-β and G_(i)-γ are mixed andused to transfect monolayers of HEK293 cells using the calcium phosphateprecipitate method (M. Wigler at al., Cell. 1977; 11; 223 and DS. Im atal., Mol. Pharmacol. 2000; 57; 753). Briefly, a DNA mixture containing25 μg of DNA and 0.25 M CaCl₂ is added to HEPES-buffered 2 mM Na₂HPO₄.Subconfluent monolayers of HEK293 cells are poisoned with 25 mMchloroquine, and the DNA precipitate is then applied to the cells. After4 h, the monolayers are washed with phosphate-buffered saline and refedmedia (90% 1:1 Dulbecco's modified essential media (DMEM):F-12+10% fetalbovine serum). The cells are harvested 48-72 h after addition of the DNAby scraping in HME buffer (in mM: 20 HEPES, 5 MgCl₂, 1 EDTA, pH 7.4)containing 10% sucrose on ice, and disrupted using a Dounce homogenizer.After centrifugation at 800×g, the supernatant is diluted with HMEwithout sucrose and centrifuged at 100,000×g for 1 h. The resultingpellet is rehomogenized and centrifuged a second hour at 100,000×g. Thiscrude membrane pellet is resuspended in HME with sucrose, aliquoted, andsnap-frozen by immersion in liquid nitrogen. The membranes are stored at70° C. Protein concentration is determined spectroscopically by Bradfordprotein assay.

GTPγS Binding Assay Using S1P Receptor/HEK293 Membrane Preparations

GTPγS binding experiments are performed as described by DS. Im et al.,Mol. Pharmacol. 2000; 57:753. Ligand-mediated GTPγS binding toG-proteins is measured in GTP binding buffer (in mM: 50 HEPES, 100 NaCl,10 MgCl₂, pH 7.5) using 25 μg of a membrane preparation from transientlytransfected HEK293 cells. Ligand is added to membranes in the presenceof 10 μM GDP and 0.1 nM [³⁵S]GTPγS (1200 Ci/mmol) and incubated at 30°C. for 30 min. Bound GTPγS is separated from unbound using the Brandelharvester (Gaithersburg, Md.) and counted with a liquid scintillationcounter.

The composition of the invention preferably contains 0.01 to 20% byweight of S1P receptor agonists, more preferably 0.1 to 10%, e.g. 0.5 to5% by weight, based on the total weight of the composition.

The sugar alcohol may act as a diluent, carrier, filler or bulkingagent, and may suitably be mannitol, maltitol, inositol, xylitol orlactitol, preferably a substantially non-hygroscopic sugar alcohol, e.g.mannitol (D-mannitol). A single sugar alcohol may be used, or a mixtureof two or more sugar alcohols, e.g a mixture of mannitol and xylitol,e.g. in a ratio of 1:1 to 4:1.

In a particularly preferred embodiment, the sugar alcohol is preparedfrom a spray-dried composition, e.g. mannitol composition, having a highspecific surface area. The use of this type of mannitol composition mayassist in promoting uniform distribution of the S1P receptor agonistthroughout the mannitol in the composition. A higher surface area may beachieved by providing a sugar alcohol, e.g. mannitol, preparationconsisting of particles having a smaller mean size and/or a roughersurface on each particle. The use of a spray-dried sugar alcohol, e.g.mannitol, e.g. with a mean particle size of 300 μm or less, has alsobeen found to improve compressibility and hardness of tablets formedfrom the composition.

Preferably the single point surface area of the sugar alcoholpreparation, e.g. mannitol, is 1 to 7 m²/g, e.g. 2 to 6 m²/g or 3 to 5m²/g. The mannitol preparation may suitably have a mean particle size of100 to 300 μm, e.g. 150 to 250 μm and a bulk density of 0.4 to 0.6 g/mL,e.g. 0.45 to 0.55 g/mL. A suitable high surface area mannitol is ParteckM200, available commercially from E. Merck.

The composition preferably contains 75 to 99.99% by weight of the sugaralcohol, more preferably 85 to 99.9%, e.g 90 to 99.5% by weight, basedon the total weight of the composition.

The composition preferably further comprises a lubricant. Suitablelubricants include stearic acid, magnesium stearate, calcium stearate,zinc stearate, glyceryl palmitostearate, sodium stearyl fumarate, canolaoil, hydrogenated vegetable oil such as hydrogenated castor oil (e.g.Cutina® or Lubriwax® 101), mineral oil, sodium lauryl sulfate, magnesiumoxide, colloidal silicon dioxide, silicone fluid, polyethylene glycol,polyvinyl alcohol, sodium benzoate, talc, poloxamer, or a mixture of anyof the above. Preferably the lubricant comprises magnesium stearate,hydrogenated castor oil or mineral oil. Colloidal silicon dioxide andpolyethylene glycol are less preferred as the lubricant.

The composition preferably contains 0.01 to 5% by weight of thelubricant, more preferably 1 to 3% by weight, e.g. about 2% by weight,based on the total weight of the composition.

The composition may comprise one or more further excipients such ascarriers, binders or diluents. In particular, the composition maycomprise microcrystalline cellulose (e.g. Avicel®), methylcellulose,hydroxypropylcellulose, hydroxypropylmethylcellulose, starch (e.g. cornstarch) or dicalcium phosphate, preferably in an amount of from 0.1 to90% by weight, e.g. 1 to 30% by weight, based on the total weight of thecomposition. Where a binder, e.g. microcrystalline cellulose,methylcellulose, hydroxypropyl cellulose, hydroxypropylmethyl celluloseis used, it is preferably included in an amount of 1 to 8%, e.g. 3 to 6%by weight, based on the total weight of the composition. The use of abinder increases the granule strength of the formulation, which isparticularly important for fine granulations. Microcrystalline celluloseand methylcellulose are particularly preferred where a high tablethardness and/or longer disintegration time is required. Hydroxypropylcellulose is preferred where faster disintegration is required. Whereappropriate, xylitol may also be added as an additional binder, forexample in addition to microcrystalline cellulose, e.g. in an amount upto 20% by weight of the sugar alcohol, e.g. xylitol.

In one embodiment, the composition further comprises a stabiliser,preferably glycine HCl or sodium bicarbonate. The stabiliser may bepresent in an amount of e.g. 0.1 to 30%, preferably 1 to 20% by weight.

The composition may be in the form of a powder, granule or pellets or aunit dosage form, for example as a tablet or capsule. The compositionsof the present invention are well-adapted for encapsulation into anorally administrable capsule shell, particularly a hard gelatin shell.

Alternatively the compositions may be compacted into tablets. Thetablets may optionally be coated, for instance with talc or apolysaccharide (e.g. cellulose) or hydroxypropyl methylcellulosecoating.

Where the pharmaceutical capsule is in unit dosage form, each unitdosage will suitably contain 0.5 to 10 mg of the S1P receptor agonist.

The compositions of the invention may show good stabilitycharacteristics as indicated by standard stability trials, for examplehaving a shelf life stability of up to one, two or three years, and evenlonger. Stability characteristics may be determined, e.g. by measuringdecomposition products by HPLC analysis after storage for particulartimes, at particular temperatures, e.g. 20°, 40° or 60° C.

The pharmaceutical compositions of the present invention may be producedby standard processes, for instance by conventional mixing, granulating,sugar-coating, dissolving or lyophilizing processes. Procedures whichmay be used are known in the art, e.g. those described in L. Lachman etal. The Theory and Practice of Industrial Pharmacy, 3rd Ed, 1986, H.Sucker et al, Pharmazeutische Technologie, Thieme, 1991, Hagers Handbuchder pharmazeutischen Praxis, 4th Ed. (Springer Verlag, 1971) andRemington's Pharmaceutical Sciences, 13th Ed., (Mack Publ., Co., 1970)or later editions.

In one aspect, the present invention relates to a process for producinga pharmaceutical composition, comprising:

(a) mixing an S1P receptor agonist with a sugar alcohol;(b) milling and/or granulating the mixture obtained in (a); and(c) mixing the milled and/or granulated mixture obtained in (b) with alubricant.

By using this process, a preparation having a good level of content andblend uniformity (i.e. a substantially uniform distribution of the S1Preceptor agonist throughout the composition), dissolution time andstability is obtained.

The S1P receptor agonist, e.g.2-amino-2-[2-(4-octylphenyl)ethyl]propane-1,3-diol, hydrochloride, mayoptionally be micronized, and/or pre-screened, e.g. with a 400 to 500 μmmesh screen, before step (a) in order to remove lumps. The mixing step(a) may suitably comprise blending the S1P receptor agonist and thesugar alcohol, e.g. mannitol in any suitable blender or mixer for e.g.100 to 400 revolutions.

The process may be carried out by dry mixing the components. In thisembodiment the milling step (b) may suitably comprise passing themixture obtained in (a) through a screen, which preferably has a meshsize of 400 to 500 μm. Process step (a) may comprise the step of mixingthe total amount of S1P receptor agonist at first with a low amount ofsugar alcohol, e.g. from 5 to 25% by weight of the total weight of sugaralcohol, in order to form a pre-mix. Subsequently the remaining amountof sugar alcohol is added to the pre-mix. Step (a) may also comprise thestep of adding a binder solution, e.g. methylcellulose and/or xylitol,e.g. an aqueous solution, to the mixture. Alternatively the binder isadded to the mix dry and water is added in the granulation step.

The milled mixture obtained in (b) may optionally be blended once morebefore mixing with the lubricant. The lubricant, e.g. magnesiumstearate, is preferably pre-screened, e.g. with a 800 to 900 μm screen,before mixing.

Alternatively, a wet granulation process is employed. In thisembodiment, the S1P receptor agonist is preferably first dry-mixed withthe desired sugar alcohol, e.g. mannitol, and the obtained sugaralcohol/S1P receptor agonist mixture is then dry-mixed with a bindersuch as hydroxypropyl cellulose or hydroxypropylmethyl cellulose. Wateris then added and the mixture granulated, e.g. using an automatedgranulator. The granulation is then dried and milled.

If desirable, an additional amount of binder may be added in step (c) tothe mixture obtained in (b).

The process may comprise a further step of tabletting or encapsulatingthe mixture obtained in (c), e.g. into a hard gelatin capsule using anautomated encapsulation device. The capsules may be coloured or markedso as to impart an individual appearance and to make them instantlyrecognizable. The use of dyes can serve to enhance the appearance aswell as to identify the capsules. Dyes suitable for use in pharmacytypically include carotenoids, iron oxides, and chlorophyll. Preferably,the capsules of the invention are marked using a code.

The pharmaceutical compositions of the present invention are useful,either alone or in combination with other active agents, for thetreatment and prevention of conditions e.g. as disclosed in U.S. Pat.No. 5,604,229, WO 97/24112, WO 01/01978, U.S. Pat. No. 6,004,565, U.S.Pat. No. 6,274,629 and JP-14316985, the contents of which areincorporated herein by reference.

In particular, the pharmaceutical compositions are useful for:

a) treatment and prevention of organ or tissue transplant rejection, forexample for the treatment of the recipients of heart, lung, combinedheart-lung, liver, kidney, pancreatic, skin or corneal transplants, andthe prevention of graft-versus-host disease, such as sometimes occursfollowing bone marrow transplantation; particularly in the treatment ofacute or chronic silo- and xenograft rejection or in the transplantationof insulin producing cells, e.g. pancreatic islet cells;b) treatment and prevention of autoimmune disease or of inflammatoryconditions, e.g. multiple sclerosis, arthritis (for example rheumatoidarthritis), inflammatory bowel disease, hepatitis, etc.;c) treatment and prevention of viral myocarditis and viral diseasescaused by viral myocarditis, including hepatitis and AIDS.

Accordingly, in further aspects the present invention provides:

1. A composition as defined above, for use in treating or preventing adisease or condition as defined above.2. A method of treating a subject in need of immunomodulation,comprising administering to the subject an effective amount of acomposition as defined above.3. A method of treating or preventing a disease or condition as definedabove, comprising administering to the subject a composition as definedabove.4. Use of a pharmaceutical composition as defined above for thepreparation of a medicament for the prevention or treatment of a diseaseor condition as defined above.

The invention will now be described with reference to the followingspecific embodiments.

EXAMPLE 1

Micronized Compound A, e.g.2-amino-2-[2-(4-octylphenyl)ethyl]propane-1,3-diol, hydrochloride salt(FTY720), is screened and 116.7 g of the screened compound is mixed with9683.3 g mannitol (Parteck M200 from E. Merck). The mixture is thenmilled in a Frewitt MGI device (Key International Inc. USA) using a 30mesh screen. Magnesium stearate is screened using a 20 mesh screen and200 g of the screened compound blended with the FTY720/mannitol mixtureto produce a product composition.

The product composition is then compacted on a tablet press using a 7 mmdie to form 120 mg tablets, each containing:

Compound A, e.g. FTY720 * 1.4 mg Mannitol M200 116.2 mg Magnesiumstearate 2.4 mg Total 120 mg * 1 mg of Compound A in free form isequivalent to 1.12 mg of FTY720.

EXAMPLE 2

In a further example, the process of example 1 is repeated except thatthe magnesium stearate is replaced by Cutina® (hydrogenated castor oil).

EXAMPLE 3

Compound A, e.g. FTY720, and mannitol (Parteck M200 from E. Merck) areeach screened separately using an 18 mesh screen. 1.9 g screened FTY720is mixed with 40 g screened mannitol for 120 revolutions in a blender at32 rpm. The FTY720/mannitol mixture is then screened through a 35 meshscreen.

The screened FTY720/mannitol mixture is added to a granulator along witha further 340.1 g mannitol and 12 g hydroxypropylcellulose. The mixtureis mixed for 3 minutes. Water is then added at a rate of 100 ml/minuteand the mixture granulated for 2 minutes. The granulation is transferredinto a tray dryer and dried at 50° C. for 150 minutes.

The mixture is then milled in a Frewitt MGI device using a 35 meshscreen. Magnesium stearate is screened and 6 g of the screened compoundis blended for 90 revolutions at 32 rpm with the FTY720/mannitol mixtureto produce a product composition showing a substantially uniformdistribution of the S1P receptor agonist throughout the mannitol in theblend.

The product composition is then filled into size 3 hard gelatin shellson an Hoflinger & Karg 400 encapsulation device. 120 mg of the productcomposition is added to each capsule. Therefore each capsule contains:

FTY720 * 0.56 mg Mannitol M200 114.04 mg Hydroxypropylcellulose 3.6 mgMagnesium stearate 1.8 mg Total 120 mg

EXAMPLE 4

In a further example, the process of example 3 is repeated except thatthe magnesium stearate is replaced by Cutina® (hydrogenated castor oil).

EXAMPLE 5

In a further example, the process of example 3 is repeated except thatthe hydroxypropyl cellulose is replaced by hydroxypropylmethylcellulose.

EXAMPLE 6a

Micronized Compound A, e.g. FTY720, is screened using a 400 μm (40 mesh)screen. 58.35 g of the screened compound is mixed with 4841.65 gmannitol (Parteck M200 from E. Merck) in a 25 L Bohle bin blender for240 blending revolutions. The mixture is then milled in a Frewitt MGIdevice using a 400 μm mesh screen, and the milled mixture is blendedonce more. Magnesium stearate is screened and 100 g of the screenedcompound is blended with the FTY720/mannitol mixture to produce aproduct composition showing a substantially uniform distribution of theS1P receptor agonist throughout the mannitol in the blend.

The product composition is then filled into size 3 hard gelatin shellson an Hoflinger & Karg 400 encapsulation device. 120 mg of the productcomposition is added to each capsule. Therefore each capsule contains:

FTY720 * 1.4 mg Mannitol M200 116.2 mg Magnesium stearate 2.4 mg Total120 mg

EXAMPLE 6b

In an alternative embodiment, capsules are manufactured using thecomponents and in the amounts as described in Example 6a, but the FTY720is first mixed with 14 mg mannitol (before screening). This mixture isthen screened as described above. The screened mixture is then blendedwith the remaining mannitol and the magnesium stearate is added,followed by additional blending and filling into capsules.

EXAMPLES 7 AND 8

In further examples, capsules are prepared as described in example 6,except that each capsule contains each component in the followingamounts:

Example 7 Example 8 FTY720 * 2.8 mg 5.6 mg Mannitol M200 114.8 mg 112 mgMagnesium stearate 2.4 mg 2.4 mg Total 120 mg 120 mg

EXAMPLES 9 TO 11

In further examples, capsules are prepared as described in examples 6 to8, except that the magnesium stearate is replaced in each case byCutina® (hydrogenated castor oil).

EXAMPLES 12 TO 22

In further examples, capsules or tablets are prepared as described inexamples 1 to 11, except that FTY720 is replaced in each case by2-amino-2-{2-[4-(1-oxo-5-phenylpentyl)phenyl]ethyl}propane-1,3-diolhydrochloride.

EXAMPLES 23 AND 24

Capsules containing the following ingredients are prepared, by weighingeach component and mixing in a mortar, then filling into capsules:

Example 23 Example 24 FTY720 5 mg 1 mg D-mannitol 83.7 mg 117 mg Cornstarch 24 mg — Avicel ® PH101 12 mg — Hydroxypropylcellulose 0.3 mg 7 mgTalc 3 mg 3 mg Lubri wax ®101 2 mg 2 mg Total 130 mg 130 mg

EXAMPLES 25 TO 27

Pharmaceutical compositions containing the following ingredients areproduced:

Example 25 Example 26 Example 27 FTY720 5 g 10 g 100 g D-mannitol 991 g986 g 897 g Methylcellulose SM-25 4 g 4 g 3 g Total 1000 g 1000 g 1000 g

The FTY720 and a proportion of the D-mannitol equal to twice the weightof the FTY720 are mixed in a Microspeed Mixer MS-5 type (Palmer, USA)for 2 minutes at 1200 rpm. The remaining D-mannitol is added to themixture and mixed for another 2 minutes. 80 or 60 milliliters of 5%methylcellulose SM-25 solution is supplied from a hopper and granulatedunder the same conditions. The mixture is extruded through a screen with0.4 mm apertures using an extruder RG-5 type. The extruded material isdried at 65° C. by a fluidized-bed granulator STREA I Type (Patheon,Canada) and then sieved through a 24 mesh sieve. Fine particles whichpass through a 60 mesh sieve are removed. The obtained fine granules arefilled into capsules by a Zuma capsule-filling machine (100 mg percapsule).

EXAMPLES 28 TO 31

Tablets, containing the following ingredients (in mg) are produced:

Example 28 Example 29 Example 30 Example 31 FTY720 1 1 1 1 D-mannitol62.3 62.3 62.0 62.0 Xylitol* 26.7(5.4) 26.7(5.4) 26.6 26.6 Methyl- — —0.4 0.4 cellulose Microcrystal- 24.0 — 24.0 — line cellulose Low- — 24.0— 24.0 substituted Hydroxy- propyl- cellulose Hydrogenated 6.0 6.0 6.06.0 oil Total 120.0 120.0 120.0 120.0 *The amount of xylitol indicatedin brackets was used as a binder.

FTY720, D-mannitol and xylitol are placed in a fluid-bed granulator(MP-01 model, Powrex), mixed for five minutes, and granulated underspray of binder solution, followed by drying till the exhausttemperature reaches 40° C. The granulation conditions are as shownbelow. Dried powder is passed through a 24-mesh sieve, added to thespecified amount of filler and lubricant, and mixed in a mixer (TubularMixer, WAB) for three minutes to make the powder for compression.

The resulting powder is compressed by a tabletting machine (Cleanpresscorrect 12 HUK, Kikushui Seisakusho) with a punch of 7 mm i.d.×7.5 mm Rat a compression force of 9800 N.

Granulation Conditions:

Item Setting Charge-in amount 1170 g Volume of intake-air 50 m³/minTemperature of intake-air 75° C. Flow rate of spray solution 15 mL/minSpray air pressure 15 N/cm² Spray air volume 30 L/min Volume of bindersolution 351 mL

EXAMPLES 32 TO 39

Tablets containing the following ingredients (in mg) are produced:

Ex. 32 Ex. 33 Ex. 34 Ex. 35 Ex. 36 Ex. 37 Ex. 38 Ex. 39 FTY720 1 1 1 1 11 1 1 D-mannitol 116.6 114.2 104.6 114.2 104.6 116.6 115.4 113 magnesium2.4 2.4 2.4 2.4 2.4 — — — stearate glycine HCl — 2.4 12 — — — — — sodium— — — 2.4 12 — — — bicarbonate zinc — — — — — 2.4 — — stearate siliconefluid — — — — — — 3.6 — mineral oil — — — — — — — 6 Total 120.0 120.0120.0 120.0 120.0 120.0 120.0 120.0

1. A solid pharmaceutical composition suitable for oral administration,comprising: (a) a S1P receptor agonist; and (b) a sugar alcohol.
 2. Acomposition according to claim 1, wherein the S1P receptor agonistcomprises 2-amino-2-[2-(4-octylphenyl)ethyl]propane-1,3-diol or2-amino-2-{2-[4-(1-oxo-5-phenylpentyl)-phenyl]ethyl}propane-1,3-diol ora pharmaceutically acceptable salt thereof.
 3. A composition accordingto claim 1 or claim 2, wherein the sugar alcohol comprises mannitol. 4.A composition according to any preceding claim, further comprising alubricant.
 5. A composition according to claim 4, wherein the lubricantcomprises magnesium stearate.
 6. A composition according to anypreceding claim, comprising 0.5 to 5% by weight of the S1P receptoragonist.
 7. A composition according to any preceding claim, comprising90 to 99.5% by weight of the sugar alcohol.
 8. A composition accordingto any of claims 4 to 7, comprising 1.5 to 2.5% by weight of thelubricant.
 9. A composition according to any preceding claim, in theform of a tablet.
 10. A composition according to any of claims 1 to 9 inthe form of a capsule.